CN101357142A - Use of Clostridium butyricum in preparing medicine composition for preventing and treating cerebrovascular disease - Google Patents
Use of Clostridium butyricum in preparing medicine composition for preventing and treating cerebrovascular disease Download PDFInfo
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Abstract
The invention discloses the application of a clostridium butyricum in preparing a medicament combination for preventing and treating cardiovascular and cerebrovascular diseases, wherein, the medicament combination comprises drugs, health care products and drinks, and the clostridium butyricum is selected from clostridium butyricum with the conservation number being 0313.1 in CGMCC, but the selection is not limited. The cardiovascular and cerebrovascular diseases include hyperlipidemia, hypertension, cerebral thrombosis, cerebral embolism, cerebral ischemia, myocardial infarction, coronary heart disease, cerebral infarction, cerebral hemorrhage, angina, myocarditis, etc.
Description
Technical field
The present invention relates to the new medicine use of clostridium butyricum, be specifically related to clostridium butyricum, belong to biological medicine field as the application of active ingredient in preparation prevention and treatment cardiovascular and cerebrovascular diseases medicament compositions.
Background technology
Cardiovascular and cerebrovascular disease is to threaten one of the most serious disease of human health at present, and its M ﹠ M surpasses neoplastic disease and leaps to the first in the world.The data that health ministry was announced in 2005, inland of China just had a people to die from the disease that causes because of cardiovascular and cerebrovascular vessel in average per 15 seconds.Cardiovascular and cerebrovascular disease belongs to chronic disease, needs long-term prescription, therefore, and effectively and the little treatment cardiovascular and cerebrovascular diseases medicament of toxic and side effects presses for.
Studies show that atherosclerosis (AS) is the main pathological basis of cardiovascular and cerebrovascular disease morbidity, and hyperlipidemia is atherosclerotic main inducing.Therefore, blood lipid-lowering medicine has certain curative effect to prevention and treatment cardiovascular and cerebrovascular disease.Along with the continuous development of Protocols in Molecular Biology, find that now struvite cytokine and free radical are the key factors that cardiovascular and cerebrovascular disease takes place and develops.Tumor necrosis factor (TNF-α), interleukin 8 (IL-8), interleukin-11 β struvite cytokines such as (IL-1 β) have promoted the generation and the development of atherosclerosis and cardiovascular and cerebrovascular disease, and struvite cytokine levels such as TNF-α, the IL-8 among the cardiovascular and cerebrovascular disease patients serum, IL-1 β are apparently higher than healthy people.The morbidity cause of cardiovascular and cerebrovascular disease is also relevant with free radical participation tissue injury; its product is malonaldehyde lipid peroxides (LPO) such as (MDA); it participates in inflammatory reaction; cause the artery sclerosis; even cause cell death; body has a series of protection enzymes for the defence radical damage; as superoxide dismutase (SOD); glutathion peroxidase (GSH-PX); catalase (CAT) [Liu Zhiting; pay army, Zhang Lianzhi, Wang Peng. cardiovascular and cerebrovascular disease patients serum LPO and SOD; GSH-PX; the research of CAT monitoring and dependency. Jilin medical science 200122 (1): 21-22].
So far, do not see the report of clostridium butyricum treatment cardiovascular and cerebrovascular disease as yet, the inventor finds that through further investigation clostridium butyricum has significant prevention and treatment by blood fat reducing, adjusting cytokine-expressing and elimination radical pair cardiovascular and cerebrovascular disease, and without any toxic and side effects, can prolonged application, special this patent of invention of application.
Summary of the invention
The present invention provides clostridium butyricum in preparation prevention and the application for the treatment of in the cardiovascular and cerebrovascular diseases medicament compositions first, and wherein said pharmaceutical composition comprises medicine, health product and beverage etc.
The inventor confirms that after deliberation clostridium butyricum has significant prevention, therapeutical effect to cardiovascular and cerebrovascular disease.Described cardiovascular and cerebrovascular disease comprises hyperlipemia, hypertension, cerebral thrombosis, cerebral embolism, cerebral ischemia, myocardial infarction, coronary heart disease, cerebral infarction, cerebral hemorrhage, angina pectoris, myocarditis etc.; Described prevention, therapeutical effect be meant the prevention cardiovascular and cerebrovascular disease generation and development, treatment and auxiliary treatment cardiovascular and cerebrovascular disease.
At present, studies confirm that free radical raises and struvite cytokine rising atherosclerotic generation of promotion and development in blood fat rising, the blood, thereby cause the generation and the development of cardiovascular and cerebrovascular disease.Free radical raises and struvite cytokine rising coup injury blood vessel in perhaps blood fat rising, the blood, causes the generation and the development of various cardiovascular and cerebrovascular diseases.Especially, development along with Protocols in Molecular Biology, find that cardiovascular and cerebrovascular disease also is an inflammation immunologic derangement disease, hyperlipemia, hypertension, cerebral thrombosis, cerebral embolism, cerebral ischemia, myocardial infarction, coronary heart disease, cerebral infarction, cerebral hemorrhage, angina pectoris, TNF-α in the cardiovascular and cerebrovascular disease patient blood such as myocarditis, IL-8, struvite cytokines such as IL-1 β generally raise, struvite cytokine has promoted the generation and the development of cardiovascular and cerebrovascular disease, TNF-α, IL-8, struvite cytokines such as IL-1 β have been considered to the important indicator of cardiovascular and cerebrovascular disease observation of curative effect.
The inventor studies show that clostridium butyricum not only can blood fat reducing and eliminate free radical in the blood, also the balance to human cytokine has very strong adjusting function, can regulate the unconventionality expression of cytokine, make it recover normal level, the damage of sexual cell factor pair blood vessel that diminishes inflammation, thus reach effective prevention and treatment to cardiovascular and cerebrovascular disease.The inventor studies confirm that clostridium butyricum through fermentation culture, can produce the butyric acid (butanoic acid) of 40mmol/L.The regulating action of the clostridium butyricum pair cell factor realizes that by entering a large amount of butyric acid of intestinal secretion butyric acid mainly by regulating the expression of nuclear factor (NF-κ B), is eliminated dysimmunity.The regulatory function of the pair cell factor is the clostridium butyricum functions peculiar, and probiotic bacterias such as bacillus bifidus, lactobacillus are not owing to produce butyric acid, short-chain fatty acid such as their excretory lactic acid, acetic acid also must be converted into the regulating action of the butyric acid competence exertion pair cell factor, so can not directly eliminate of the damage of the struvite cytokine of unconventionality expression, thereby reach effective prevention and treatment to cardiovascular and cerebrovascular disease to blood vessel.
The inventor studies show that intestinal immune disorder and enteritis, and not only the struvite cytokine of intestinal mucosa raises, and the struvite cytokine in the blood also significantly raises.Struvite cytokine in the blood raises and will cause or promote the generation and the development of cardiovascular and cerebrovascular disease.Therefore, enteritis is the risk factor that cardiovascular and cerebrovascular disease takes place and develops, the inventor studies confirm that struvite cytokines such as TNF-α, IL-8 that clostridium butyricum can be by the overexpression effectively eliminate enteritis time of secretion butyric acid, IL-1 β, effectively prevention, treatment cardiovascular and cerebrovascular disease.
Clostridium butyricum is the result who plays a role from many-sides such as blood lipid regulation, cytokine, elimination free radicals to the treatment of cardiovascular and cerebrovascular disease, but most critical and main be to regulate cytokine expression, the damage of sexual cell factor pair blood vessel diminishes inflammation, improve interleukin-11 0 anti-inflammatory cytokines levels such as (IL-10), thereby reach effective prevention and treatment cardiovascular and cerebrovascular disease.The present invention has disclosed the novel medical use of clostridium butyricum adjusting cytokine therapy cardiovascular and cerebrovascular disease first, and its effectiveness has all obtained confirmation by pharmacodynamic study and clinical observation, does not see relevant report.Clostridium butyricum is expected to play a significant role in the generation of treatment cardiovascular and cerebrovascular disease and prevention cardiovascular and cerebrovascular disease and development, and clostridium butyricum is without any toxic and side effects, can take for a long time, its application dose also is that the clinician is easy to grasp and adjust according to the state of an illness.
Clostridium butyricum of the present invention is selected from but is not limited to clostridium butyricum (Clostridium butyricum), CGMCC, preservation numbering 0313.1.
Clostridium butyricum of the present invention refers to the living organism individuality.
The present invention is that clostridium butyricum with effective dose is according to top described as medicament active composition, according to certain preparation process, add excipient substances such as conventional excipient, flavoring agent, disintegrating agent, antiseptic, lubricant, wetting agent, adhesive, solvent, thickening agent, solubilizing agent, make any dosage form that is suitable for using clinically, as dosage forms such as tablet, capsule, granule, powder, liquid preparation, enemas.
Clostridium butyricum compositions of the present invention can be that clostridium butyricum is made active bacteria formulation as active ingredient separately, also can be that active bacteria formulation is made in the combination of clostridium butyricum and other active ingredients, to play synergistic therapeutic action, improve prevention and therapeutic effect to cardiovascular and cerebrovascular disease.
Indication effective dose of the present invention is meant with clostridium butyricum can not be lower than 1 * 10 according to total viable count that the top described solid live bacteria preparation of making as medicament active composition alone or in combination comprises
6CFU/g is generally 1 * 10
7More than the CFU/g, can reach 1 * 10
12CFU/g or 1 * 10
12More than the CFU/g.
Indication effective dose of the present invention is meant with clostridium butyricum can not be lower than 1 * 10 according to total viable count that the top described liquid active bacteria formulation of making as medicament active composition alone or in combination comprises
6CFU/mL is generally 1 * 10
7More than the CFU/mL, can reach 1 * 10
12CFU/mL or 1 * 10
12More than the CFU/mL.
The preferred preparation process of the present invention is, inoculates strain in the first liquid medium within, cultivates or multistage amplification cultivation, and is then that liquid culture is centrifugal, collects wet bacterium mud, and with described bacterium mud drying, sieve, obtain dry mycopowder.The dry mycopowder of gained is mixed with pharmaceutical carrier, make final dosage form.Preparation process is not limited to of the present invention, and other known preparation process is all passable.
Because it is the application of medicament active composition in preparation prevention, treatment cardiovascular and cerebrovascular diseases medicament with the clostridium butyricum that the present invention discloses first; therefore; make medicament as medicament active composition and adjuvant combination alone or in combination with clostridium butyricum; so long as this medicament is used for prevention, treatment cardiovascular and cerebrovascular disease, all belong to protection scope of the present invention.
Clostridium butyricum of the present invention all has the effect of prevention, treatment cardiovascular and cerebrovascular disease when making any dosage form.Any medicament; prepare patent medicine if contain the clostridium butyricum composition in its component; on signs such as its packing or description or on other any propaganda materials,, then fall within protection scope of the present invention as long as indicate or point out the effect that has prevention, treats cardiovascular and cerebrovascular disease.
Clostridium butyricum of the present invention can be made health product or beverage.Health product or beverage with clostridium butyricum is made indicate or point out the effect that has prevention, treats cardiovascular and cerebrovascular disease if need only on signs such as its packing or description or on other any propaganda materials, then fall within protection scope of the present invention.
The specific embodiment
The explanation of medication preparation example: the preparation of clostridium butyricum preparation is at first to prepare mycopowder, doses corresponding adjuvant then as required and makes final dosage form according to a conventional method.Used culture medium in the mycopowder preparation process, the present technique field known culture medium of personnel is all passable.The preparation of clostridium butyricum preparation of the present invention is all passable according to preparation process well known by persons skilled in the art, be not limited to method described below, the present invention specifically is prepared as example with oral clostridium butyricum viable capsule, the preparation method of clostridium butyricum preparation is described, those skilled in the art are according to the preparation process that is easy to grasp other dosage form of the present invention, at this is interest of clarity, and the preparation method of other dosage form is omitted, narration explanation no longer one by one.
The preparation of the oral clostridium butyricum viable capsule of medication preparation embodiment
The preparation of 1 mycopowder
Get clostridium butyricum strain Guan Yizhi, being dissolved in sterilizes is equipped with in the 100mL triangular flask of 10mL normal saline and an amount of bead, 75 ℃ of water-baths activate 10 minutes, draw the 1mL bacteria suspension with the 1mL aseptic straw, inoculation is equipped with in the 250mL triangular flask of 50mL amplification culture medium, putting the interior 37 ℃ of constant temperature vibrations of rocking bed (190rpm) cultivated 24 hours, switching is equipped with in the 450mL amplification culture medium 2500mL plate washer triangular flask, 37 ℃ of constant-temperature shaking culture 24 hours, microscopy is not transferred in the seed tank that the 4.5L amplification culture medium is housed after having assorted bacterium again, anaerobism was cultivated (aeration quantity 3: 1) 24 hours, microscopy does not have to transfer behind the assorted bacterium and has in the fermentation tank of 45L fermentation medium, 37 ℃ of anaerobism were cultivated (aeration quantity 3: 1) 24 hours, and microscopy spore rate reaches more than 80%, stopped to cultivate.Use continuous centrifuge, 12000rpm is centrifugal.Collect wet bacterium mud, weigh, add freeze drying protectant by 1: 1 (w/v), lyophilizing, porphyrize is crossed 100 mesh sieves, and room temperature is preserved standby.
The preparation of 2 finished products
According to the viable count of clostridium butyricum mycopowder, add defatted milk powder, glucose in proportion, make viable count be not less than 1 * 10
6CFU/g, encapsulated.
Test example 1 clostridium butyricum is to the regulating action research of atherosclerotic rat cytokine and blood fat
1 materials and methods
1.1 the male Sprague-Dawley of laboratory animal, medicine and reagent (SD) rat, 180-220g, available from dimension tonneau China Experimental Animal Center, the cleaning level, the quality certification number is: SCXP (capital) (2002)-0003.The clostridium butyricum mycopowder is produced by Qingdao DongHai Pharmacy Co., Ltd.Rat TNF-α, IL-1 β, IL-10 ELISA test kit.
1.2 rat AS model set up rat at the lumbar injection vitamin D
3(behind 600,000 units/Kg), give high lipid food (3% cholesterol, 0.5% sodium cholate, 0.2% propylthiouracil, 5% white sugar, 10% Adeps Sus domestica, 81.3% basic feedstuff) and fed for 8 weeks, can form typical A S pathological changes.
1.3 the reagent preparation is diluted to 10 with the clostridium butyricum mycopowder
6CFU/mL.
1.4 grouping and medication are divided into 3 groups at random with rat, i.e. normal control group, model control group and medication therapy groups, 6 every group.Except that normal control group feed normal feedstuff, all the other each groups are all at the lumbar injection vitamin D
3After give high lipid food, simultaneously medication therapy groups gives clostridium butyricum (10
6CFU/mL) irritate stomach, each 1mL/100g (body weight), 1 time/day.Normal control group and model control group give the water with volume.After 8 weeks, the lumbar injection chloral hydrate anesthesia is got aortic blood and is carried out index determining.
1.5 index determining
1.5.1TNF-the ratio that α, IL-1 β, IL-10 measure in the 4mL/g tissue adds normal saline, uses glass homogenizer homogenate, draws supernatant behind the centrifugal 10min of homogenate 3000r/min, adopts the ELISA method to measure, and is undertaken by the test kit explanation.
1.5.2 measure the serum lipid parameter according to a conventional method, comprising: T-CHOL (TC), triglyceride (TG) and HDL-C (HDL-C).
2 results
2.1TNF-α, IL-1 β, IL-10 and normal control group are relatively, all significantly risings (P＜0.01) of TNF-α, IL-1 β, IL-10 level in the model control group tremulous pulse, clostridium butyricum can significantly reduce TNF-α in the tremulous pulse, IL-1 β level, significantly increase IL-10 level (P＜0.01), see Table 1.
Table 1 clostridium butyricum is to the influence of AS rat TNF-α, IL-1 β, IL-10
2.2 serum lipid parameter model control group TC, TG are significantly higher than normal control group (P＜0.05), model control group HDL-C significantly is lower than normal control group (P＜0.05); Clostridium butyricum treatment back is compared TC, TG with model control group and is significantly reduced (P＜0.05), and HDL-C significantly raises (P＜0.05), sees Table 2.
Table 2 clostridium butyricum is to the influence of AS rat TC, TG, HDL-C
3 discuss
AS is the pathologic basis of cardiovascular and cerebrovascular disease, AS is similar to other diseases associated with inflammation such as rheumatoid arthritis, psoriasis, asthma and inflammatory bowel disease, with the inflammation immunoreaction abnormity, struvite cytokine overexpressions such as TNF-α, IL-8, IL-1 β, quicken generation and the development of AS, and then quickened the generation and the development of cardiovascular and cerebrovascular disease.
Studies show that struvite cytokine levels such as cardiovascular and cerebrovascular disease patient TNF-α, IL-1 β such as hyperlipemia, hypertension, cerebral thrombosis, cerebral embolism, cerebral ischemia, myocardial infarction, coronary heart disease, cerebral infarction, cerebral hemorrhage, angina pectoris, myocarditis all significantly raise.TNF-α as an apoptotic intracellular signal can the induction of vascular endothelial apoptosis, promote the generation of endothelin level (ET-1) and increase the weight of the damage of blood vessel wall, and the excretory effect of promotion interleukin 6 (IL-6) is arranged, may command liposoluble enzymatic activity also, suppress lipoprotein lipase (LPL) activity, suppress the generation of the local LPL of speckle, thereby be unfavorable for the lipid dissolving, and be easy to be deposited on blood vessel wall, promote arteriosclerosis to form, cause the generation of various cardiovascular and cerebrovascular diseases.TNF-α can bring out myocarditis, lowers cardiac function and causes myocardium cell necrosis by necrosis or apoptosis, causes angina pectoris and arrhythmia etc.The positive detection rate of cardiovascular and cerebrovascular disease patient IL-1 β is along with the also corresponding rising of the rising of TNF-α verification and measurement ratio, two kinds of cytokines of TNF-α, IL-1p are by reciprocal induction, interaction, fellowship the pathophysiological process of cardiovascular and cerebrovascular diseases such as hyperlipemia, hypertension, cerebral thrombosis, cerebral embolism, cerebral ischemia, myocardial infarction, coronary heart disease, cerebral infarction, cerebral hemorrhage, angina pectoris, myocarditis, thereby cause the generation and the development of cardiovascular and cerebrovascular disease.
IL-10 is a kind of anti-inflammatory cytokines with strong immunoregulation effect, brings into play protective effect in cardiovascular and cerebrovascular disease.Clostridium butyricum can make unusual cytokine-expressing recover normal or improve, and to stoping struvite cytokine the infringement of cardiovascular and cerebrovascular vessel and cell is played a good role, and can effectively prevent, treat cardiovascular and cerebrovascular disease.Clostridium butyricum mainly is by secretion butyric acid pair cell factor performance regulating action.
Blood fat mainly is meant cholesterol and the triglyceride in the serum.No matter be that cholesterol level increases, or the content of triglyceride increases, or both all increase, and are referred to as hyperlipemia.Hyperlipemia is the key factor that atherosclerosis, cardiovascular and cerebrovascular disease and microcirculation disturbance take place.Clinical data shows that the incidence rate of cardiovascular and cerebrovascular disease increases with the concentration rising of serum cholesterol and triglyceride.It is the apodeictic fact that blood fat reducing can obviously reduce coronary heart disease danger, and serum cholesterol reduces by 1%, and coronary heart disease is dangerous to reduce by 2%.Therefore, the discovery early of hyperlipemia and active treatment have very important meaning to the control of cardiovascular and cerebrovascular disease.One of function of high density lipoprotein (HDL) is that transportation endogenous cholesterol to liver is handled, so study of anti-atherogenic effect is arranged.In inspection, reflect the level of HDL indirectly by the content of checking HDL-C.HDL-C reduces during atherosclerosis, shows that HDL reduces.Clostridium butyricum reaches the effect of blood fat reducing by reducing TC, TG and rising HDL, thereby effectively prevents and treat atherosclerosis, cardiovascular and cerebrovascular disease and microcirculation disturbance.Clostridium butyricum awaits further investigation to TC, TG, HDL-C regulating action mechanism.
The regulating action research of cytokine unconventionality expression in the blood when testing example 2 clostridium butyricums to enteritis
1 experiment material and method
1.1 the male Sprague-Dawley of laboratory animal, medicine and reagent (SD) rat, 180-220g, available from dimension tonneau China Experimental Animal Center, the cleaning level, the quality certification number is: SCXP (capital) (2002)-0003.The clostridium butyricum mycopowder is produced by Qingdao DongHai Pharmacy Co., Ltd.Freund's complete adjuvant, IL-8ELISA test kit, TNF-α ELISA test kit.The anti-Mus NF-of rabbit κ Bp65 monoclonal antibody, S-P immunohistochemical staining test kit.
1.2 experimental technique
1.2.1 the new calves colon is got in the preparation of cattle mucous membrane of colon albumen lyophilized powder, scrapes and gets the cattle mucous membrane of colon, freezes molten method repeatedly and obtains cattle mucous membrane of colon albumen, purification concentrates, and is standby in 4 ℃ of preservations behind the vacuum freeze-drying.
1.2.2 cattle mucous membrane of colon albumen is got in the foundation of enteritis rat model and complete Freund's adjuvant (1: 1) is made complete antigen, select for use body weight the SD of 200 ± 20g rat, the modeling rat is every interior injections of antigens 4mg of the sufficient sole of the foot first, in the 10th, 17,24,31 day respectively at the sufficient sole of the foot, back, groin, intraperitoneal injection antigen 6mg, last 1 injection does not add adjuvant, reaches necessarily to serum IgG and tires.
1.2.3 the reagent preparation is diluted to 10 with the clostridium butyricum mycopowder
6CFU/mL.
1.2.4 normal control group (8) is established in animal grouping and treatment.Modeling success back (there is secretions on the intestinal mucosa surface, epithelial cell shedding, inflammatory cell infiltration, row's mucus loose stool) is with modeling rat random packet, 1) model control group (10), give normal saline and irritate stomach; 2) clostridium butyricum (10
6CFU/mL) treatment group (10).Each 1mL/100g body weight, 1 time/day, irritate stomach, give normal saline or curative 21 days continuously.1.2.5IL-8, TNF-α index determining gives normal saline or medicine to the after 21 days, the sacrificed by decapitation rat is got blood, 4 ℃ of standing over night, 3000rpm, 20min is centrifugal, gets supernatant, packing, be put in-20 ℃ to be measured.Use the ELISA test kit, to specifications, the content of rat IL-8 and TNF-α in the mensuration rat blood serum.1.2.6NF-kB activity is measured and is got colonic pathological change position tissue, makes immunohistochemical staining.It is positive that cell is pale brown color, be positioned nucleus and (or) Cytoplasm.Epithelial cell positive findings computational methods: count 50 crypts of far-end colon (from anus about 4cm upwards) at random, count the positive percentage of each crypts, and mark by following standard :≤5% is 0 minute; 6%～25% is 1 minute, note+; 26%～50% is 2 minutes, note ++; 51%～75% is 3 minutes, note +++; 76%～100% is 4 minutes, note ++ ++.Calculate the average of every rat afterwards.
1.2.7 t-student ' t check is used in the significance,statistical check of the data difference between each group of statistical procedures, significance of difference boundary is P＜0.05.
2 experimental results
2.1 serum il-8 content
IL-8 content in the model control group rat blood serum is than the remarkable rising (P＜0.05) of normal control group, the treatment of clostridium butyricum treatment group rat after 21 days the IL-8 content in the serum compare remarkable reduction (P＜0.05) with model control group, see Table 1.
Table 1 clostridium butyricum is to the influence of enteritis rat blood serum IL-8 changes of contents
2.2 serum TNF-alpha content
TNF-alpha content and normal control group in the model control group rat blood serum significantly raises (P＜0.05), clostridium butyricum treatment group rat treatment after 21 days the TNF-alpha content in the serum compare remarkable reduction (P＜0.05) with model control group, see Table 2.
The influence that table 2 clostridium butyricum changes enteritis rat blood serum TNF-alpha content
2.3NF-kB activity
Model control group rat NF-κ B expresses with the normal control group and significantly raises (P＜0.05), the treatment of clostridium butyricum treatment group rat after 21 days NF-κ B expression compare remarkable reduction (P＜0.05) with model control group, see Table 3.
The influence that table 3 clostridium butyricum is expressed enteritis rat NF-κ B
3 discuss
The method of setting up enteritis is a lot, this selection be to set up the enteritis model by the method for autoimmune disorder, be equivalent to ulcerative colitis, observe the regulating action that struvite cytokine that clostridium butyricum causes enteritis raises.Studies show that intestinal tract diseases such as chronic diarrhea, irritable bowel syndrome, ulcerative colitis all have inflammation performance in various degree, struvite cytokine-expressing levels such as the IL-8 in intestinal mucosa and the blood, TNF-α all raise.Struvite cytokine in the blood raises and must cause damage to the heart and cerebrovascular and tissue, promotes the generation and the development of cardiovascular and cerebrovascular disease.At present, studies confirm that butyric acid (butanoic acid) can be by regulating NF-kB activity, regulating cell factor expression.Originally discover that IL-8, TNF-alpha content are accompanied by the decline of NF-κ B expression and reduce after the clostridium butyricum treatment, show that clostridium butyricum can suppress the NF-kB activity by secretion butyric acid and regulate struvite cytokine expression such as IL-8, TNF-α in the enteritis patient blood, make it recover normal level, thereby prevent, treat cardiovascular and cerebrovascular disease.
Test example 3 clostridium butyricums are eliminated free radical research in the blood
1 materials and methods
1.1 laboratory animal, reagent and instrument cleaning level kunming mice, male and female half and half available from Qingdao City medicine inspecting institute, are fed and are observed test after 3 days.The clostridium butyricum mycopowder is produced by Qingdao DongHai Pharmacy Co., Ltd; Superoxide dismutase (SOD), malonaldehyde (MDA), glutathion peroxidase (GSH-PX) test kit are purchased and are built up bio-engineering research institute in Nanjing.Visible spectrophotometer; Water-bath; Tissue mashing machine; Desk centrifuge; Scalpel; Operating scissors.
1.2 method
1.2.1 30 of mices of grouping are divided into 3 groups at random, i.e. normal control group, model control group, clostridium butyricum treatment group, and 10 every group with the male Mus sub-cage rearing of female Mus.Duration of test food and water supply capacity.
1.2.2 moulding and administration
1.2.2.1 the reagent preparation is diluted to 10 with the clostridium butyricum mycopowder
6CFU/mL.
1.2.2.2 irritating stomach, moulding and administration clostridium butyricum or sterile saline carry out twice of every day every day at duration of test; Moulding once a day, is total to 6 weeks of moulding beginning in the 4th day.Each dosage and number of times be the normal control group 1.: the injecting normal saline 0.18mL/ of nape portion of every day, sterile saline is only irritated stomach 0.2mL/ after 0.5 hour.2. model control group: the D-galactose normal saline solution 150mg/ (kg days) of nape portion subcutaneous injection 2%, sterile saline is only irritated stomach 0.2mL/ after 0.5 hour.3. clostridium butyricum group: every day nape portion subcutaneous injection 2% D-galactose normal saline solution 150mg/ (kg days), after 0.5 hour with 10
6CFU/mL clostridium butyricum mycopowder diluent is only irritated stomach 0.2mL/.Finish after modeling and the medication and put to death white mice, get brain, the heart tissue of mice respectively, clean in the ice bath that to be prepared into tissue homogenate cold preservation standby with dislocation method.
1.3 the mensuration of index with above-mentioned homogenate 3500r/min centrifugal after, get supernatant and measure SOD, GSH-PX, MDA in brain, the heart tissue respectively according to the method for test kit description.
1.4 date processing adopts t check and X
2Date processing is carried out in check, and significance,statistical is P＜0.05 or P＜0.01.
2 results
By table 1,2,3 as can be known: the content of the SOD in model control group mouse brain, the heart tissue, GSH-PX and MDA has been compared significant difference with the normal control group, and the modeling success is described; Clostridium butyricum can improve SOD, the GSH-PX activity in model mice brain and the heart significantly, and the utmost point reduces the MDA level of brain and heart tissue significantly.
Table 1 is respectively organized the comparison of SOD, MDA, GSH-PX in the mouse brain tissue
Annotate: with the contrast of normal control group, * P＜0.01; With the model control group contrast, ▲ ▲ P＜0.01
Table 2 is respectively organized the comparison of SOD, MDA, GSH-PX in the mouse heart tissue
Annotate: with the contrast of normal control group, * P＜0.01; With the model control group contrast, ▲ ▲ P＜0.01
3 discuss
The morbidity cause of cardiovascular and cerebrovascular disease is relevant with free radical participation tissue injury, and its product is malonaldehyde lipid peroxides (LPO) such as (MDA), and it has participated in inflammatory reaction, and vascular endothelial cell injury causes the artery sclerosis, even causes cell death.Body has a series of protection enzymes, as superoxide dismutase (SOD), glutathion peroxidase (GSH-PX) for the defence radical damage.
MDA is the final metabolite of lipid peroxidation, and its content has reflected the level and the level of lipid peroxidation of free radical.The main component SOD of free radical resisting and GSH-PX have then reflected the ability of body removing free radical in the body.Free radical not only can directly cause the lipid peroxidation injury of cell membrane, also can generate MDA, forms MDA-LDL with low density lipoprotein, LDL (LDL) and causes the target cell damage.Free-radical generating too much still can stimulate inflammatory cell to discharge more cytokine and increase the synthetic of leukotriene, causes, increases the weight of vascular endothelial injury, promotes atherosclerosis and thrombosis.This result of study prompting clostridium butyricum has free radical resisting enzymatic activitys such as the SOD of raising and GSH-PX, removes free radical, and protection cardiovascular and cerebrovascular vessel and tissue prevent and the treatment cardiovascular and cerebrovascular disease takes place and the effect of development.Clostridium butyricum awaits further investigation to superoxide dismutase (SOD), glutathion peroxidase (GSH-PX), malonaldehyde (MDA) regulating action mechanism.
The present invention in implementation process employed microorganism fungus kind in (No. 13, North No.1 Row, Zhongguancun, Haidian District, Beijing City, Institute of Microorganism, Academia Sinica's postcode 100080) preservation on July 28th, 1997 at China Committee for Culture Collection of Microorganisms common micro-organisms center.But clostridium butyricum of the present invention is not limited to the microorganism fungus kind of following preservation.
(1) classification name: clostridium butyricum Clostridium butyricum, preserve numbering 0313.1.
The mentioned microorganism strain detects through this microorganism center, and testing result is survival.
Claims (9)
1, the application of clostridium butyricum in preparation prevention, treatment cardiovascular and cerebrovascular diseases medicament compositions.
2,, it is characterized by clostridium butyricum and refer to bion alive by the described application of claim 1.
3, by the described application of claim 1, it is characterized by clostridium butyricum including but not limited to clostridium butyricum CGMCC0313.1 strain.
4, by the described application of claim 1, it is characterized by cardiovascular and cerebrovascular disease and comprise hyperlipemia, hypertension, cerebral thrombosis, cerebral embolism, cerebral ischemia, myocardial infarction, coronary heart disease, cerebral infarction, cerebral hemorrhage, angina pectoris, myocarditis.
5,, it is characterized by the application in preparation prevention, the cardiovascular and cerebrovascular diseases caused pharmaceutical composition of the struvite cytokine of treatment by the described application of claim 1.
6,, it is characterized by raise application in the cardiovascular and cerebrovascular diseases caused pharmaceutical composition of preparation prevention, treatment blood fat by the described application of claim 1.
7,, it is characterized by the application in preparation prevention, the cardiovascular and cerebrovascular diseases caused pharmaceutical composition of treatment free radical by the described application of claim 1.
8, by the described application of claim 1, it is characterized by the clostridium butyricum compositions can be that clostridium butyricum is made active bacteria formulation as active ingredient separately, also can be that active bacteria formulation is made in the combination of clostridium butyricum and other active ingredients.
9, by the described application of claim 1, it is characterized by pharmaceutical composition and comprise medicine, health product, beverage.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2007101375419A CN101357142B (en) | 2007-08-05 | 2007-08-05 | Use of Clostridium butyricum in preparing medicine composition for preventing and treating cerebrovascular disease |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2007101375419A CN101357142B (en) | 2007-08-05 | 2007-08-05 | Use of Clostridium butyricum in preparing medicine composition for preventing and treating cerebrovascular disease |
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| Publication Number | Publication Date |
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| CN101357142A true CN101357142A (en) | 2009-02-04 |
| CN101357142B CN101357142B (en) | 2012-08-22 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105012349A (en) * | 2015-08-06 | 2015-11-04 | 温州医科大学 | Probiotic preparation for preventing and treating vascular dementia and preparation method of probiotic preparation |
| CN105147740A (en) * | 2015-08-06 | 2015-12-16 | 浙江省台州医院 | Pharmaceutical composition for improving cerebral ischemic injury as well as preparation method and application of pharmaceutical composition |
| CN110652528A (en) * | 2010-06-04 | 2020-01-07 | 国立大学法人东京大学 | Composition for inducing proliferation or accumulation of regulatory T cells |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101095698B (en) * | 2006-06-26 | 2010-12-01 | 青岛东海药业有限公司 | Use of Clostridium Butyricum for preventing and treating foetid faeces toxin syndrome and disease |
-
2007
- 2007-08-05 CN CN2007101375419A patent/CN101357142B/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110652528A (en) * | 2010-06-04 | 2020-01-07 | 国立大学法人东京大学 | Composition for inducing proliferation or accumulation of regulatory T cells |
| CN105012349A (en) * | 2015-08-06 | 2015-11-04 | 温州医科大学 | Probiotic preparation for preventing and treating vascular dementia and preparation method of probiotic preparation |
| CN105147740A (en) * | 2015-08-06 | 2015-12-16 | 浙江省台州医院 | Pharmaceutical composition for improving cerebral ischemic injury as well as preparation method and application of pharmaceutical composition |
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| Publication number | Publication date |
|---|---|
| CN101357142B (en) | 2012-08-22 |
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Effective date of registration: 20091204 Address after: No. 8, Shanghai Road, Jiaonan, Qingdao, Shandong Applicant after: Qingdao Donghai Pharmaceutical Co., Ltd. Co-applicant after: Beijing Puerkang Medical High Technology Co., Ltd. Address before: No. 8, Shanghai Road, Jiaonan, Qingdao, Shandong Applicant before: Qingdao Donghai Pharmaceutical Co., Ltd. Co-applicant before: Dongfanghaixin Biological Technology Co., Ltdd., Beijing |
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